FIG. 8.

Planar fluorescence reflectance imaging analysis of crimped TEVV constructs. TEVVs were seeded with CFSE-labeled endothelial cells (ECs) that stained positive for vWF (a; green=vWF, blue=DAPI, red=phalloidin; b=isotype control). After seeding (stereomicroscopy analysis of the ECs on the leaflet; c, d; scale bar=500 μm) the valves were crimped (for 10 min, ∼5 mm) and the fluorescence signal was measured before and after crimping (e; V1–V3 show three representative examples). When quantified, the fluorescence in the seeded constructs decreased during the crimping procedure, while the fluorescence signal of unseeded control areas remained unchanged (f). Interestingly, the cellular loss significantly decreased when increased static conditioning periods were used, indicating that adapted protocols may prevent the EC loss (f) (scale bars: a, 16 μm; b, 25 μm). Color images available online at www.liebertpub.com/tec